The Derivatives of Cromolyn Ameliorate the Abnormal Misfolding of Amyloid Proteins and Neuroinflammation in the Neural Cells.

BACKGROUND: The representative symptom of Alzheimer's Disease (AD) has mainly been mentioned to be misfolding of amyloid proteins, such as amyloid-beta (Aß) and tau protein. In addition, the neurological pathology related to neuroinflammatory signaling has recently been raised as an important feature in AD. Currently, numerous drug candidates continue to be investigated to reduce symptoms of AD, including amyloid proteins misfolding and neuroinflammation. OBJECTIVE: Our research aimed to identify the anti-AD effects of two chemical derivatives modified from cromoglicic acid, CNU 010 and CNU 011. METHODS: CNU 010 and CNU 011 derived from cromoglicic acid were synthesized. The inhibitory effects of Aß and tau were identified by thioflavin T assay. Moreover, western blots were conducted with derivates CNU 010 and CNU 011 to confirm the effects on inflammation. RESULTS: CNU 010 and CNU 011 significantly inhibited the aggregation of Aß and tau proteins. Moreover, they reduced the expression levels of mitogen-activated protein (MAP) kinase and nuclear factor kappa-light-chain-enhancer of activated B cells (NF- κB) signaling proteins, which are representative early inflammatory signaling markers. Also, the inhibitory effects on the lipopolysaccharide (LPS)-induced cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) expression referring to late inflammation were confirmed. CONCLUSION: Our results showing multiple beneficial effects of cromolyn derivatives against abnormal aggregation of amyloid proteins and neuroinflammatory signaling provide evidence that CNU 010 and CNU 011 could be further developed as potential drug candidates for AD treatment.

High-density EEG of auditory steady-state responses during stimulation of basal forebrain parvalbumin neurons.

We present high-density EEG datasets of auditory steady-state responses (ASSRs) recorded from the cortex of freely moving mice with or without optogenetic stimulation of basal forebrain parvalbumin (BF-PV) neurons, known as a subcortical hub circuit for the global workspace. The dataset of ASSRs without BF-PV stimulation (dataset 1) contains raw 36-channel EEG epochs of ASSRs elicited by 10, 20, 30, 40, and 50 Hz click trains and time stamps of stimulations. The dataset of ASSRs with BF-PV stimulation (dataset 2) contains raw 36-channel EEG epochs of 40-Hz ASSRs during BF-PV stimulation with latencies of 0, 6.25, 12.5, and 18.75 ms and time stamps of stimulations. We provide the datasets and step-by-step tutorial analysis scripts written in Python, allowing for descriptions of the event-related potentials, spectrograms, and the topography of power. We complement this experimental dataset with simulation results using a time-dependent perturbation on coupled oscillators. This publicly available dataset will be beneficial to the experimental and computational neuroscientists.

Distinct subnetworks of the thalamic reticular nucleus.

The thalamic reticular nucleus (TRN), the major source of thalamic inhibition, regulates thalamocortical interactions that are critical for sensory processing, attention and cognition1-5. TRN dysfunction has been linked to sensory abnormality, attention deficit and sleep disturbance across multiple neurodevelopmental disorders6-9. However, little is known about the organizational principles that underlie its divergent functions. Here we performed an integrative study linking single-cell molecular and electrophysiological features of the mouse TRN to connectivity and systems-level function. We found that cellular heterogeneity in the TRN is characterized by a transcriptomic gradient of two negatively correlated gene-expression profiles, each containing hundreds of genes. Neurons in the extremes of this transcriptomic gradient express mutually exclusive markers, exhibit core or shell-like anatomical structure and have distinct electrophysiological properties. The two TRN subpopulations make differential connections with the functionally distinct first-order and higher-order thalamic nuclei to form molecularly defined TRN-thalamus subnetworks. Selective perturbation of the two subnetworks in vivo revealed their differential role in regulating sleep. In sum, our study provides a comprehensive atlas of TRN neurons at single-cell resolution and links molecularly defined subnetworks to the functional organization of thalamocortical circuits.

Paraspinal back muscles in asymptomatic volunteers: quantitative and qualitative analysis using computed tomography (CT) and magnetic resonance imaging (MRI).

BACKGROUND: To evaluate paraspinal back muscles of asymptomatic subjects using qualitative and quantitative analysis on CT and MRI and correlate the results with demographic data. METHODS: Twenty-nine asymptomatic subjects were enrolled prospectively (age: mean 34.31, range 23-50; 14 men, 15 women) from August 2016 to April 2017. Qualitative analysis of muscles was done using Goutallier's system on CT and MRI. Quantitative analysis entailed cross sectional area (CSA) on CT and MRI, Hounsfield unit (HU) on CT, fat fraction using two-point Dixon technique on MRI. Three readers independently analyzed the images; intra- and inter-observer agreements were measured. Linear regression and Spearman's analyses were used for correlation with demographic data. RESULTS: CSA values were significantly higher in men (p < 0.001). Fat fraction was higher (22.53% vs. 14.35%) and HU lower (36.00 vs. 47.43) in women (p < 0.001). Intra- and inter-observer reliabilities of the two methods were greater than 0.8, except for CSA of L5/S1 on MRI; however, regarding quantitative analysis, decreasing HU and increasing fat fraction were correlated with increasing age, female gender and lower lumbar segment (p < 0.001). CONCLUSION: MRI and CT can be reliably used for qualitative and quantitative analysis of paraspinal back muscles, regarding fat content. Fat fraction and HU showed highest reliabilities.

Differential modulation of NREM sleep regulation and EEG topography by chronic sleep restriction in mice.

Compensatory elevation in NREM sleep EEG delta power has been typically observed following prolonged wakefulness and widely used as a sleep homeostasis indicator. However, recent evidence in human and rodent chronic sleep restriction (CSR) studies suggests that NREM delta power is not progressively increased despite of accumulated sleep loss over days. In addition, there has been little progress in understanding how sleep EEG in different brain regions responds to CSR. Using novel high-density EEG electrode arrays in the mouse model of CSR where mice underwent 18-h sleep deprivation per day for 5 consecutive days, we performed an extensive analysis of topographical NREM sleep EEG responses to the CSR condition, including period-amplitude analysis of individual slow waves. As previously reported in our analysis of REM sleep responses, we found different patterns of changes: (i) progressive decrease in NREM sleep duration and consolidation, (ii) persistent enhancement in NREM delta power especially in the frontal and parietal regions, and (iii) progressive increases in individual slow wave slope and frontal fast oscillation power. These results suggest that multiple sleep-wake regulatory systems exist in a brain region-specific manner, which can be modulated independently, especially in the CSR condition.

Distinct Topographical Patterns of Spike-Wave Discharge in Transgenic and Pharmacologically Induced Absence Seizure Models.

Absence seizures (AS) are generalized non-convulsive seizures characterized by a brief loss of consciousness and spike-and-wave discharges (SWD) in an electroencephalogram (EEG). A number of animal models have been developed to explain the mechanisms of AS, and thalamo-cortical networks are considered to be involved. However, the cortical foci have not been well described in mouse models of AS. This study aims to use a high density EEG in pathophysiologically different AS models to compare the spatiotemporal patterns of SWDs. We used two AS models: a pharmacologically induced model (gamma-hydroxybutyric acid, GHB model) and a transgenic model (phospholipase beta4 knock-out, PLCß4 model). The occurrences of SWDs were confirmed by thalamic recordings. The topographical analysis of SWDs showed that the onset and propagation patterns were markedly distinguishable between the two models. In the PLCß4 model, the foci were located within the somatosensory cortex followed by propagation to the frontal cortex, whereas in the GHB model, a majority of SWDs was initiated in the prefrontal cortex followed by propagation to the posterior cortex. In addition, in the GHB model, foci were also observed in other cortical areas. This observation indicates that different cortical networks are involved in the generation of SWDs across the two models.

Optogenetic stimulation of basal forebrain parvalbumin neurons modulates the cortical topography of auditory steady-state responses.

High-density electroencephalographic (hdEEG) recordings are widely used in human studies to determine spatio-temporal patterns of cortical electrical activity. How these patterns of activity are modulated by subcortical arousal systems is poorly understood. Here, we couple selective optogenetic stimulation of a defined subcortical cell-type, basal forebrain (BF) parvalbumin (PV) neurons, with hdEEG recordings in mice (Opto-hdEEG). Stimulation of BF PV projection neurons preferentially generated time-locked gamma oscillations in frontal cortices. BF PV gamma-frequency stimulation potently modulated an auditory sensory paradigm used to probe cortical function in neuropsychiatric disorders, the auditory steady-state response (ASSR). Phase-locked excitation of BF PV neurons in advance of 40 Hz auditory stimuli enhanced the power, precision and reliability of cortical responses, and the relationship between responses in frontal and auditory cortices. Furthermore, synchronization within a frontal hub and long-range cortical interactions were enhanced. Thus, phasic discharge of BF PV neurons changes cortical processing in a manner reminiscent of global workspace models of attention and consciousness.

Desynchronization of Theta Oscillations in Prefrontal Cortex during Self-stimulation of the Medial Forebrain Bundles in Mice.

Stimulation of the medial forebrain bundle (MFB) can reinforce intracranial self-stimulation (ICSS) in rodents (i.e., reward-seeking behavior). The MFB stimulation produces a highly reliable behavioral output that enabled a clear distinction of the animal behavioral states between the non-ICSS and ICSS periods. However, the cortical states during these reward-seeking behaviors are not fully characterized in comparison to those during volitional behavior. This study was designed to characterize the cortical rhythms of and coherence between prefrontal cortex and hippocampus during the wheel-turning behavior reinforced by the ICSS in comparison to the wheel-turning without ICSS. We used a wheel for freely moving mice, which was programmed to deliver cathode currents through an electrode in the MFB at each one-quarter turn of the wheel to induce ICSS. The wheel-turning epochs were extracted from the pre-ICSS, ICSS and post-ICSS sessions and the prefrontal EEGs and the hippocampal LFPs in the epochs were analyzed with power and synchronization analyses. During the ICSS, the EEG power decreased at 6~10 Hz in the prefrontal cortex, while was not significantly altered in the hippocampus. Furthermore, we found that the phase synchrony between the prefrontal cortex and the hippocampus corresponding to information transmission between the two regions during reward-seeking motion decreased preceding MFB stimulation reinforced by ICSS. Our findings suggest that theta-activity can be reliably dissociated from active behavior if the animal is involved in self-stimulation.

A missense allele of KARRIKIN-INSENSITIVE2 impairs ligand-binding and downstream signaling in Arabidopsis thaliana.

A smoke-derived compound, karrikin (KAR), and an endogenous but as yet unidentified KARRIKIN INSENSITIVE2 (KAI2) ligand (KL) have been identified as chemical cues in higher plants that impact on multiple aspects of growth and development. Genetic screening of light-signaling mutants in Arabidopsis thaliana has identified a mutant designated as ply2 (pleiotropic long hypocotyl2) that has pleiotropic light-response defects. In this study, we used positional cloning to identify the molecular lesion of ply2 as a missense mutation of KAI2/HYPOSENSITIVE TO LIGHT, which causes a single amino acid substitution, Ala219Val. Physiological analysis and genetic epistasis analysis with the KL-signaling components MORE AXILLARY GROWTH2 (MAX2) and SUPPRESSOR OF MAX2 1 suggested that the pleiotropic phenotypes of the ply2 mutant can be ascribed to a defect in KL-signaling. Molecular and biochemical analyses revealed that the mutant KAI2ply2 protein is impaired in its ligand-binding activity. In support of this conclusion, X-ray crystallography studies suggested that the KAI2ply2 mutation not only results in a narrowed entrance gate for the ligand but also alters the structural flexibility of the helical lid domains. We discuss the structural implications of the Ala219 residue with regard to ligand-specific binding and signaling of KAI2, together with potential functions of KL-signaling in the context of the light-regulatory network in Arabidopsis thaliana.

Pulse-train Stimulation of Primary Somatosensory Cortex Blocks Pain Perception in Tail Clip Test.

Human studies of brain stimulation have demonstrated modulatory effects on the perception of pain. However, whether the primary somatosensory cortical activity is associated with antinociceptive responses remains unknown. Therefore, we examined the antinociceptive effects of neuronal activity evoked by optogenetic stimulation of primary somatosensory cortex. Optogenetic transgenic mice were subjected to continuous or pulse-train optogenetic stimulation of the primary somatosensory cortex at frequencies of 15, 30, and 40 Hz, during a tail clip test. Reaction time was measured using a digital high-speed video camera. Pulse-train optogenetic stimulation of primary somatosensory cortex showed a delayed pain response with respect to a tail clip, whereas no significant change in reaction time was observed with continuous stimulation. In response to the pulse-train stimulation, video monitoring and local field potential recording revealed associated paw movement and sensorimotor rhythms, respectively. Our results show that optogenetic stimulation of primary somatosensory cortex at beta and gamma frequencies blocks transmission of pain signals in tail clip test.

Gamma-Band Activities in Mouse Frontal and Visual Cortex Induced by Coherent Dot Motion.

A key question within systems neuroscience is to understand how the brain encodes spatially and temporally distributed local features and binds these together into one perceptual representation. Previous works in animal and human have shown that changes in neural synchrony occur during the perceptual processing and these changes are distinguished by the emergence of gamma-band oscillations (GBO, 30-80 Hz, centered at 40 Hz). Here, we used the mouse electroencephalogram to investigate how different cortical areas play roles in perceptual processing by assessing their GBO patterns during the visual presentation of coherently/incoherently moving random-dot kinematogram and static dots display. Our results revealed that GBO in the visual cortex were strongly modulated by the moving dots regardless of the existence of a global dot coherence, whereas GBO in frontal cortex were modulated by coherence of the motion. Moreover, concurrent GBO across the multiple cortical area occur more frequently for coherently moving dots. Taken together, these findings of GBO in the mouse frontal and visual cortex are related to the perceptual binding of local features into a globally-coherent representation, suggesting the dynamic interplay across the local/distributed networks of GBO in the global processing of optic flow.

Differential modulation of global and local neural oscillations in REM sleep by homeostatic sleep regulation.

Homeostatic rebound in rapid eye movement (REM) sleep normally occurs after acute sleep deprivation, but REM sleep rebound settles on a persistently elevated level despite continued accumulation of REM sleep debt during chronic sleep restriction (CSR). Using high-density EEG in mice, we studied how this pattern of global regulation is implemented in cortical regions with different functions and network architectures. We found that across all areas, slow oscillations repeated the behavioral pattern of persistent enhancement during CSR, whereas high-frequency oscillations showed progressive increases. This pattern followed a common rule despite marked topographic differences. The findings suggest that REM sleep slow oscillations may translate top-down homeostatic control to widely separated brain regions whereas fast oscillations synchronizing local neuronal ensembles escape this global command. These patterns of EEG oscillation changes are interpreted to reconcile two prevailing theories of the function of sleep, synaptic homeostasis and sleep dependent memory consolidation.

Changes in effective connectivity of sensorimotor rhythms in thalamocortical circuits during the induction and recovery of anesthesia in mice.

The thalamocortical network serves a role in both consciousness and sensorimotor processing. However, little is known regarding how changes in conscious states, via induction of and recovery from anesthesia, affect the processing of sensorimotor information in the thalamocortical network. To address this, we investigated the dynamics of causal interactions among sensorimotor rhythms (SMR; frequency range of 3-12Hz) across the thalamocortical network during transitions into and out of ketamine-induced unconsciousness. Two local field potentials from the ventral lateral and ventrobasal thalamic nuclei, as well as two intracranial electroencephalography signals from the primary sensory and primary motor regions, were recorded in 10 mice. Spectral Granger causality analysis revealed two distinct frequency-specific patterns in sensorimotor rhythms. For the low-frequency (3-6.5Hz) SMR, loss of consciousness evoked causal influences directed from the cortex to the thalamus. For the high-frequency (6.5-12Hz) SMR, causal influences from the primary sensory cortex to other regions during the conscious period were abruptly altered by loss of consciousness and gradually regenerated following recovery of consciousness. The results of the present study indicate that anesthesia alters the flow of sensorimotor information in the thalamocortical network and may provide evidence of the neural basis of loss and recovery of sensorimotor function associated with anesthesia.

Pathophysiological implication of CaV3.1 T-type Ca2+ channels in trigeminal neuropathic pain.

A crucial pathophysiological issue concerning central neuropathic pain is the modification of sensory processing by abnormally increased low-frequency brain rhythms. Here we explore the molecular mechanisms responsible for such abnormal rhythmicity and its relation to neuropathic pain syndrome. Toward this aim, we investigated the behavioral and electrophysiological consequences of trigeminal neuropathic pain following infraorbital nerve ligations in CaV3.1 T-type Ca(2+) channel knockout and wild-type mice. CaV3.1 knockout mice had decreased mechanical hypersensitivity and reduced low-frequency rhythms in the primary somatosensory cortex and related thalamic nuclei than wild-type mice. Lateral inhibition of gamma rhythm in primary somatosensory cortex layer 4, reflecting intact sensory contrast, was present in knockout mice but severely impaired in wild-type mice. Moreover, cross-frequency coupling between low-frequency and gamma rhythms, which may serve in sensory processing, was pronounced in wild-type mice but not in CaV3.1 knockout mice. Our results suggest that the presence of CaV3.1 channels is a key element in the pathophysiology of trigeminal neuropathic pain.

Characterization of topographically specific sleep spindles in mice.

STUDY OBJECTIVE: Sleep spindles in humans have been classified as slow anterior and fast posterior spindles; recent findings indicate that their profiles differ according to pharmacology, pathology, and function. However, little is known about the generation mechanisms within the thalamocortical system for different types of spindles. In this study, we aim to investigate the electrophysiological behaviors of the topographically distinctive spindles within the thalamocortical system by applying high-density EEG and simultaneous thalamic LFP recordings in mice. DESIGN: 32-channel extracranial EEG and 2-channel thalamic LFP were recorded simultaneously in freely behaving mice to acquire spindles during spontaneous sleep. SUBJECTS: Hybrid F1 male mice of C57BL/6J and 129S4/svJae. MEASUREMENTS AND RESULTS: Spindle events in each channel were detected by spindle detection algorithm, and then a cluster analysis was applied to classify the topographically distinctive spindles. All sleep spindles were successfully classified into 3 groups: anterior, posterior, and global spindles. Each spindle type showed distinct thalamocortical activity patterns regarding the extent of similarity, phase synchrony, and time lags between cortical and thalamic areas during spindle oscillation. We also found that sleep slow waves were likely to associate with all types of sleep spindles, but also that the ongoing cortical decruitment/ recruitment dynamics before the onset of spindles and their relationship with spindle generation were also variable, depending on the spindle types. CONCLUSION: Topographically specific sleep spindles show distinctive thalamocortical network behaviors.

Reduction in cortical gamma synchrony during depolarized state of slow wave activity in mice.

EEG gamma band oscillations have been proposed to account for the neural synchronization crucial for perceptual integration. While increased gamma power and synchronization is generally observed during cognitive tasks performed during wake, several studies have additionally reported increased gamma power during sleep or anesthesia, raising questions about the characteristics of gamma oscillation during impaired consciousness and its role in conscious processing. Phase-amplitude modulation has been observed between slow wave activity (SWA, 0.5-4 Hz) and gamma oscillations during ketamine/xylazine anesthesia or sleep, showing increased gamma activity corresponding to the depolarized (ON) state of SWA. Here we divided gamma activity into its ON and OFF (hyperpolarized) state components based on the phase of SWA induced by ketamine/xylazine anesthesia and compared their power and synchrony with wake state levels in mice. We further investigated the state-dependent changes in both gamma power and synchrony across primary motor and primary somatosensory cortical regions and their interconnected thalamic regions throughout anesthesia and recovery. As observed previously, gamma power was as high as during wake specifically during the ON state of SWA. However, the synchrony of this gamma activity between somatosensory-motor cortical regions was significantly reduced compared to the baseline wake state. In addition, the somatosensory-motor cortical synchrony of gamma oscillations was reduced and restored in an anesthetic state-dependent manner, reflecting the changing depth of anesthesia. Our results provide evidence that during anesthesia changes in long-range information integration between cortical regions might be more critical for changes in consciousness than changes in local gamma oscillatory power.

Characterization of phase transition in the thalamocortical system during anesthesia-induced loss of consciousness.

The thalamocortical system plays a key role in the breakdown or emergence of consciousness, providing bottom-up information delivery from sensory afferents and integrating top-down intracortical and thalamocortical reciprocal signaling. A fundamental and so far unanswered question for cognitive neuroscience remains whether the thalamocortical switch for consciousness works in a discontinuous manner or not. To unveil the nature of thalamocortical system phase transition in conjunction with consciousness transition, ketamine/xylazine was administered unobtrusively to ten mice under a forced working test with motion tracker, and field potentials in the sensory and motor-related cortex and thalamic nuclei were concomitantly collected. Sensory and motor-related thalamocortical networks were found to behave continuously at anesthesia induction and emergence, as evidenced by a sigmoidal response function with respect to anesthetic concentration. Hyperpolarizing and depolarizing susceptibility diverged, and a non-discrete change of transitional probability occurred at transitional regimes, which are hallmarks of continuous phase transition. The hyperpolarization curve as a function of anesthetic concentration demonstrated a hysteresis loop, with a significantly higher anesthetic level for transition to the down state compared to transition to the up state. Together, our findings concerning the nature of phase transition in the thalamocortical system during consciousness transition further elucidate the underlying basis for the ambiguous borderlines between conscious and unconscious brains. Moreover, our novel analysis method can be applied to systematic and quantitative handling of subjective concepts in cognitive neuroscience.

Changes in the thalamocortical connectivity during anesthesia-induced transitions in consciousness.

Thalamocortical networks play an important role in information integration during consciousness. However, little is known about how the information flows between the thalamus and the cortex are affected by a loss of consciousness. To investigate this issue, we analyzed effective connectivity between the cortex and the thalamus in animals during anesthesia-induced transitions. By recording the electroencephalogram from the primary motor and the primary somatosensory cortex and by recording local field potentials from the ventral lateral and the ventrobasal thalamic nuclei, we evaluated changes in the conditional Granger causality between cortical and thalamic electrical activity as mice gradually lost consciousness from the use of anesthesia (ketamine/xylazine). The point of loss of consciousness was indicated by a moment of loss of movement that was measured using a head-mounted motion sensor. The results showed that 65% of the thalamocortical information flows were changed by anesthesia-induced loss of consciousness. Specifically, the effective connectivity between the cortex and the ventral lateral thalamus was altered such that the primary motor and the primary somatosensory cortex Granger-caused the ventral lateral thalamus before loss of consciousness whereas the ventral lateral thalamus Granger-caused the primary motor cortex and the primary somatosensory cortex after loss of consciousness. In contrast, the primary somatosensory cortex consistently Granger-caused the ventrobasal thalamus, regardless of the loss of consciousness. These results suggest how information flows change across the thalamocortical network during transitions in consciousness.

The forced walking test: a novel test for pinpointing the anesthetic-induced transition in consciousness in mouse.

In consciousness or anesthesia studies, pinpointing the precise moment of consciousness or anesthetic transition has been challenging because of the variable lag time between a treatment and its induced response. Here, we describe a novel behavioral method, a forced walking test, which pinpoints the moment of the anesthetic-induced loss of motion (LOM) without handling the animals manually. The mouse is forced to walk on a treadmill, and an anesthetic drug is administered into the peritoneum via a previously secured injection route. The physical activity and the angle of head posture are tracked using a motion sensor preinstalled on the animal's head. The moments of LOM and recovery of motion (ROM) are identified from the physical activity parameters obtained by the sensor. Comparison of our method with the conventional loss-of-righting-reflex assay showed that the time point of LOM was not significantly different between the two methods when examined with two different types of anesthetic agents, propofol and ketamine/xylazine cocktail. In addition, the electrophysiological signals simultaneously acquired in the cortex and the thalamus of the mouse during the forced walking test showed that the brain rhythms induced by ketamine/xylazine anesthesia were generated and terminated in a time-locked manner with respect to LOM and ROM, respectively. In conclusion, the forced walking test allows an objective and precise detection of anesthetic-induced LOM, as well as ROM during awakening from anesthesia, in test animals.

Estimating transition point of anesthetic-induced loss of consciousness in mice by detecting motion in response to forced movement.

To characterize transition periods of entrance to and emergence from anesthetic-induced unconsciousness in terms of thalamocortical neural activity, we devised a new method estimating a transition point of anesthetic-induced loss of consciousness. The method continuously monitors an animal's head motion in response to forced movement on treadmill and uses the motion signals as a criterion of the transition. Anesthetics were administered via previously secured intraperitoneal injection route in order not to disturb the animal's spontaneous movement. Resulting signals from the motion detector could discriminate the points of entrance into and emergence from the anesthetic-induced unconsciousness with resolution corresponding to the sampling frequency. This method makes it possible to track the anesthetic transition period continuously without contaminating EEGs and LFPs.